An automated shotgun lipidomics platform for high throughput, comprehensive, and quantitative analysis of blood plasma intact lipids.
An automated shotgun lipidomics platform for high throughput, comprehensive, and quantitative analysis of blood plasma intact lipids.
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一个自动shot弹枪脂质组学平台，用于对血浆完整脂质的高吞吐量，全面和定量分析。
DOI：
10.1002/ejlt.201500145
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发表时间：
2015-10
期刊：
影响因子：
--
通讯作者：
Sampaio JL
中科院分区：
文献类型：
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作者：
Surma MA;Herzog R;Vasilj A;Klose C;Christinat N;Morin-Rivron D;Simons K;Masoodi M;Sampaio JL
血浆由于其可获得性、简单的收集和制备以及生理状态的信息读出而在脂质组学研究中获得了主导地位。同时，由于其复杂的脂质组成受到许多因素的影响，这可能会妨碍通量和/或脂质组学覆盖范围，因此在技术上也具有挑战性。为了解决这些问题，我们开发了一个全面的，高通量的，定量的基于质谱的鸟枪脂质组学平台，用于血浆脂质分析。该技术的主要特点是（i）它是全面的，涵盖22种可定量的不同脂质类别，包括200多种脂质种类;（ii）它适合高通量，少于5分钟的采集时间允许每天完全分析200份血浆样品;（iii）它通过包括测量的每种脂质类别的内标物实现绝对定量;（iv）它是高度可重复的，实现了<10%（日内）的平均变异系数，约10%（日间），约对于大多数脂质物质，15%（研究中心间）;（v）易于转移，允许直接比较不同研究中心获得的数据。此外，我们彻底评估了不同抗凝剂和冻融循环对血液稳定的影响，以排除样品制备产生的伪影。实际应用：这种鸟枪式脂质组学平台可以在不同的实验室中实施，而不会影响重现性，允许多中心研究和实验室间比较。这种可能性与高通量，广泛的脂质组学覆盖和绝对定量相结合，是临床应用和生物标志物研究的重要方面。
Blood plasma has gained protagonism in lipidomics studies due to its availability, uncomplicated collection and preparation, and informative readout of physiological status. At the same time, it is also technically challenging to analyze due to its complex lipid composition affected by many factors, which can hamper the throughput and/or lipidomics coverage. To tackle these issues, we developed a comprehensive, high throughput, and quantitative mass spectrometry-based shotgun lipidomics platform for blood plasma lipid analyses. The main hallmarks of this technology are (i) it is comprehensive, covering 22 quantifiable different lipid classes encompassing more than 200 lipid species; (ii) it is amenable to high-throughput, with less than 5 min acquisition time allowing the complete analysis of 200 plasma samples per day; (iii) it achieves absolute quantification, by inclusion of internal standards for every lipid class measured; (iv) it is highly reproducible, achieving an average coefficient of variation of <10% (intra-day), approx. 10% (inter-day), and approx. 15% (inter-site) for most lipid species; (v) it is easily transferable allowing the direct comparison of data acquired in different sites. Moreover, we thoroughly assessed the influence of blood stabilization with different anticoagulants and freeze-thaw cycles to exclude artifacts generated by sample preparation. Practical applications: This shotgun lipidomics platform can be implemented in different laboratories without compromising reproducibility, allowing multi-site studies and inter-laboratory comparisons. This possibility combined with the high-throughput, broad lipidomic coverage and absolute quantification are important aspects for clinical applications and biomarker research.